WO2016167528A1 - Procédé de conversion directe de fibroblastes humaines en cellules souches neurales à l'aide de composés à petites molécules - Google Patents

Procédé de conversion directe de fibroblastes humaines en cellules souches neurales à l'aide de composés à petites molécules Download PDF

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WO2016167528A1
WO2016167528A1 PCT/KR2016/003819 KR2016003819W WO2016167528A1 WO 2016167528 A1 WO2016167528 A1 WO 2016167528A1 KR 2016003819 W KR2016003819 W KR 2016003819W WO 2016167528 A1 WO2016167528 A1 WO 2016167528A1
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stem cells
neural stem
cells
small molecule
disease
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PCT/KR2016/003819
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Korean (ko)
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홍성회
최경아
황인식
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고려대학교산학협력단
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Priority claimed from KR1020160044187A external-priority patent/KR101816103B1/ko
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Priority to US15/545,314 priority Critical patent/US10711245B2/en
Priority to CN201680019389.1A priority patent/CN107454913B/zh
Priority to JP2017538402A priority patent/JP6599468B2/ja
Publication of WO2016167528A1 publication Critical patent/WO2016167528A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/30—Nerves; Brain; Eyes; Corneal cells; Cerebrospinal fluid; Neuronal stem cells; Neuronal precursor cells; Glial cells; Oligodendrocytes; Schwann cells; Astroglia; Astrocytes; Choroid plexus; Spinal cord tissue

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  • the present invention relates to a method for converting human fibroblasts into neural stem cells, and more particularly, to a method for directly converting from human fibroblasts to neural stem cells using a combination of small molecule compounds without introducing an external gene and its use. .
  • Retrodifferentiated stem cells have similar properties to embryonic stem cells, but viral systems, a method commonly used to form pluripotent stem cells, can mutate by random insertion of genes. Plasmids, proteins, RNA, etc. are used to solve the problem of the virus, but new problems may occur due to low efficiency and use of oncogenes.
  • neural stem cells Due to these problems, methods for directly differentiating into neural stem cells using fibroblasts have recently been studied, and various transcription factors using a viral system are introduced to induce neural stem cells from fibroblasts. Only branched transcription factors were used to reach neural stem cells. Neural stem cells can self-renew and thus get as many cells as they want and can differentiate into neurons. In addition, neural stem cells derived from fibroblasts of each individual by direct cross-differentiation method are very useful cells for the treatment of refractory brain disease because there is no ethical problem, immune rejection and tumor formation after transplantation.
  • Cell therapy is a method of implanting and treating experimentally induced healthy cells to replace damaged cells and tissues in the human body.
  • skin cells can be used to directly convert to desired cells, and genetic manipulations can be used to replace normal genes before transplanting.
  • direct cross-differentiation methods that have been studied previously are difficult to use as a practical cell therapy because they introduce a foreign gene.
  • the present inventors have made efforts to induce fibroblasts into neural stem cells using only small molecule compounds without introducing external genes, and thus, they are capable of proliferating in sufficient amounts for transplantation and are genetically stable neural stem cells without tumor formation. It was confirmed that the preparation can be completed the present invention.
  • An object of the present invention is to provide a method for producing neural stem cells by culturing human fibroblasts in a medium containing a small molecule compound, and a cell therapeutic agent for treating brain diseases containing the prepared neural stem cells as an active ingredient.
  • the present invention comprises the steps of culturing human fibroblasts in a medium comprising thiazovivin, Valproic acid, Purmorphamine, A8301, SB431542 and CHIR99021 It provides a method for producing a neural stem cell comprising.
  • the present invention also provides a cell therapy for the treatment of brain diseases containing neural stem cells produced by the above method.
  • Figure 2 shows the results of confirming the gene expression according to the addition of 13 small molecule materials.
  • Figure 3 is a result confirming the morphological changes of the cells according to the use of four, six, eight small molecule compounds.
  • 5 is a result of confirming the expression of marker genes and proteins in the early and late stages of induced neural stem cells.
  • Figure 7 shows the results of analyzing the cell proliferation rate of the induced neural stem cells.
  • FIG. 8 shows the results of confirming epigenetic genetic changes of induced neural stem cells using bisulfite PCR analysis.
  • Figure 11 shows the results of analyzing the brain of the mouse transplanted induced neural stem cells.
  • the small molecule material was used to construct an optimal combination of small molecule materials for inducing human fibroblasts into neural stem cells, and the function of each of the various small molecule materials was confirmed.
  • This induces human neural stem cells with a culture medium consisting of a combination of small molecule materials, secures genetic stability without chromosomal abnormalities, and establishes optimized culture conditions for prolonged proliferation and maintenance of induced neural stem cells.
  • the basic characteristics of human neural stem cells and the differentiation ability of trigeminal nervous system cells and various kinds of neurons were confirmed, and induced human neural stem cells were confirmed to differentiate into trigeminal neural cells without tumor formation after transplantation.
  • some small molecule substances regulate the expression of endoderm and mesoderm-related genes, the inducibility of endoderm and mesoderm-related cells was confirmed.
  • the present invention includes the step of culturing human fibroblasts in a medium comprising thiazovivin, Valproic acid, Purmorphamine, A8301, SB431542 and CHIR99021. It relates to a method for producing neural stem cells.
  • Thiazovivin N-benzyl-2- (pyrimidin-4-ylamino) thiazole-4-carboxamide
  • Rho / ROCK Rho / ROCK signal inducing apoptosis of neurons and neural stem cells and neural stem cells. It is known to block PTEN signaling, which inhibits the proliferation of cells, and is expected to inhibit neural stem cell apoptosis, increase self-renewal and self-proliferation (Matthias Groszer, et al., Science 294: 2186, 2001). .
  • the thiazovivin is a substance that selectively inhibits ROCK (Rho-associated kinase) as an inhibitor of Rho-associated kinase (ROCK), and Y-27632 may be used in addition to thiazovivin. . Treating the thiazobibin in the medium to be included in an effective concentration, the effective concentration may be affected depending on factors well known in the art, such as the type and culture method of the medium.
  • VPN valproic acid, 2-propylpentanoic acid
  • valproic acid valproic acid, 2-propylpentanoic acid
  • promote expression of cell proliferation inhibitors and genes essential for differentiation induce differentiation of cells (cancer cells), inhibit angiogenesis, and fix cell cycle to G1 status to apoptosis of cancer cells It is known to exhibit strong cytostatic anti-cancer activity by inducing apoptosis.
  • Histone deacetylase inhibits gene transcription via pRB / E2F, and the destruction of histone acetylation is associated with a number of cancer developments, and HDAC is associated with hypoxia, mortgage ( Iii) HDAC is known to be recognized as an important regulator of cell carcinogenesis and differentiation by inhibiting the expression of cell proliferation factor and expressing it under severe environmental conditions such as cell carcinogenesis.
  • the VPA is known to cause inositol reduction, inhibit GSK-3 ⁇ , activate the ERK pathway, and stimulate PPAR activity.
  • HDAC inhibitor histone deacetylase inhibitor
  • VPA valproic acid, 2-propylpentanoic acid
  • TSA trachostatin
  • the derivative may be variously pharmaceutically acceptable.
  • Inorganic or organic salts If the treatment concentration is too low, it will be difficult to see the effect. If the concentration is too high, it will be toxic, so it is necessary to check the appropriate concentration according to the cell type.
  • purmorphamine is a purine compound, and is known to be involved in the Shh signaling system.
  • the permopamine is not particularly limited as long as it can induce the Shh signal, and various derivatives may be used.
  • 2- (1-Naphthoxy) -6- (4-morpholinoanilino) -9-cyclohexylpurin) can be purchased commercially.
  • the permophamine may be treated with a medium commonly used to induce differentiation into neural stem cell-like cells. This treatment of Shh analogs, permophamine, there is an advantage that does not need to introduce a gene to produce neural stem cells from human fibroblasts.
  • the permophamine is included in the medium at an effective concentration. Effective concentrations of permopamine may be affected by factors well known in the art, such as media type and culture method.
  • A-8301 is a TGF- ⁇ type I receptor inhibitor that binds to the TGF- ⁇ type I receptor and interferes with the normal signaling process of TGF- ⁇ type I.
  • TGF- ⁇ type I Transforming growth factor- ⁇ type I
  • TGF- ⁇ type I is a multifunctional peptide that has various functions on cell proliferation, differentiation and various cell types. It is known to play a pivotal role in the growth and differentiation of various tissues such as cell differentiation and to inhibit the proliferation of neural stem cells.
  • any TGF- ⁇ type I receptor inhibitor including SB432542 may be used, and the low molecular weight TGF- ⁇ type I receptor inhibitor A-8301 may be purchased or used on the market. Can be used to promote neural stem cell proliferation by the inhibitor treatment.
  • the TGF- ⁇ type I receptor inhibitor A-8301 is treated in the medium to be included in an effective concentration. Effective concentrations may be affected by factors well known in the art, such as media type and culture method.
  • SB431542 serves to improve chromosome stability by inducing rapid reverse differentiation with an ALK5 (Activin Receptor-Like Kinase-5) inhibitor.
  • CHIR99021 is a GSK (glycogen synthase kinase) inhibitor and is a substance targeting GSK1 / 2, an upstream molecule of GSK1 / 2 involved in GSK signaling.
  • the CHIR99021 is represented as aminopyrimidine, and all GSK inhibitors may be used in addition to CHIR99021.
  • DZNep Deazaneplanocin A
  • 5-AZA 5-AZA
  • at least one small molecule compound selected from the group consisting of PD0325901, Ascorbic acid, PS48, Forskolin and Tranylcypromine, but It is not limited.
  • 5-Azacitidine means 4-amino-1- ⁇ -D-ribofuranosyl-1,3,5-triazine-2 (1H) -one It is named (4-amino-1- ⁇ -D-ribofuranosyl-1,3,5-triazin-2 (1H) -one) and is a compound known to have a DNA demethylation action.
  • 5-azacytidine is also known as an anti-neoplastic drug that is active against leukemia, lymphoma and various solid tumors.
  • the 5-azacytidine can be obtained by synthesizing by a general chemical synthesis method known in the art, or can be purchased commercially available (eg Sigma-Aldrich (St Louis, Mo, USA)).
  • PD0325901 is one of the inhibitors of MEK / ERK signaling pathway
  • ascorbic acid is one of the water-soluble vitamins and has strong antioxidant properties.
  • phospholine acts to directly activate the catalytic subunit of dadenylate cyclase to increase the concentration of intracellular cAMP, and tranylcipromine is monoamine oxidation, an enzyme that normally degrades norepinephrine at the nerve endings. It acts to inhibit enzyme (MAO).
  • the culture medium includes all media commonly used for culturing neural stem cells.
  • the medium used for culturing generally contains a carbon source, a nitrogen source and a trace element component.
  • the medium is preferably composed of DMEM / F12, N2, B27, basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF).
  • the medium for inducing stem cell culture of the present invention can be used without limitation a basal medium known in the art.
  • the basal medium may be prepared by artificially synthesizing, or a commercially prepared medium may be used.
  • commercially prepared media include Dulbecco's Modified Eagle's Medium (DMEM), Minimal Essential Medium (MEM), Basic Medium Eagle (BME), RPMI 1640, F-10, F-12, ⁇ -MEM ( ⁇ -Minimal). essential medium), G-MEM (Glasgow's Minimal Essential Medium), and Isocove's Modified Dulbecco's Medium, but are not limited thereto.
  • DMEM Dulbecco's Modified Eagle's Medium
  • MEM Minimal Essential Medium
  • BME Basic Medium Eagle
  • RPMI 1640 F-10, F-12
  • ⁇ -MEM ⁇ -Minimal
  • essential medium G-MEM (Glasgow's Minimal Essential Medium)
  • the culture period is preferably 10 to 15 days, but is not limited thereto.
  • the step of subcultured cells and suspended culture to form a sphere is preferable to further include a step of the suspension culture after the attached spear formed culture, but is not limited thereto.
  • the suspension culture and the adhesion culture are cultured for 7 to 10 days, respectively, and the step of floating culture again after attaching the formed spheres is preferably repeated 2 to 4 times, but is not limited thereto.
  • neural stem cells have a self-replicating ability and neurons and / or glia, for example, astrocytes, oligodendrocytes and / or Schwann cells ( It is an undifferentiated cell having multi-differentiation ability to differentiate into Schwann cell).
  • Neural stem cells are differentiated into neural cells, such as neurons or glia, through the steps of neural precursor cells or glia precursor cells that produce specific nervous system cells.
  • the neural stem cells may be characterized by expressing nestin, sox1 or musashi1, astrocytes, oligodendrocytes, neurons (neurons), dopamine neurons, gaba neurons , Motor neuron and choline neurons may be characterized in that the differentiation into any one or more selected from the group consisting of, but is not limited thereto.
  • the neural stem cells may be characterized by maintaining chromosome stability, and may be characterized by maintaining an undifferentiated state of 10 passages or more, but is not limited thereto.
  • the technology of the present invention is a method of ensuring genetic stability of inducing neural stem cells from human fibroblasts without introducing an external gene, and is designed to solve a method of inducing genetic defects using existing genes.
  • human fibroblasts are directly converted to neural stem cells using only a combination of small molecule materials.
  • problems of the existing technology for example, ethical and immune rejection problems associated with embryonic stem cells, tumorigenesis problems of embryonic stem cells and dedifferentiated stem cells, It is very likely to be used as a cell therapy.
  • the production of neural stem cells directly using somatic cells of brain disease patients can be used as a new cell model for studying the pathogenesis of brain diseases due to damage or death of nerve cells.
  • Neural stem cells produced in this way are very fast and economical compared to dedifferentiated stem cells.
  • De-differentiated stem cells are more than double the time and cost required to produce dedifferentiated stem cells and to differentiate them back into neurons.
  • cross-differentiated neural stem cells do not have ethical problems because they do not need to secure neural stem cells from the fetus or the adult brain, and adult stem cells have self-renewal ability.
  • cross-differentiated neural stem cells have the advantage that they can produce as many cells as desired because there is no limit to self-renewal capacity.
  • cross-differentiated neural stem cells can be produced using the patient's own cells, there is no immune rejection reaction, patient-specific cell modeling for disease research, patient-specific toxicity evaluation of the developed drug and new drug development Can be.
  • the cross-differentiated neural stem cells of the present invention do not use an external gene, there is no possibility of tumorigenesis caused by indiscriminate insertion of an external gene, and thus, the cross-differentiated neural stem cells of the present invention will be highly applicable to cell therapy.
  • the present invention relates to a cell therapy agent for treating brain diseases containing neural stem cells produced by a method comprising culturing human fibroblasts in a medium containing a small molecule compound in another aspect.
  • the brain disease may be characterized by stroke, stroke, cerebral hemorrhage, cerebral infarction, Alzheimer's disease, dementia, Huntington's disease, Parkinson's disease, multiple sclerosis, multiple nerve atrophy, epilepsy, peak disease and Creutzfeldt-Jakob disease. It is not limited to this.
  • the present invention can be used as a clinically applicable neural stem cells for the treatment of brain diseases, and can be used as a brain disease cell model that can be directly used in pathogenesis research when producing diseased neuronal stem cells derived from patients with hereditary cerebral disease.
  • a mutated gene By replacing the mutated gene with a normal gene, it can be used as a cell therapy for the treatment of patients with hereditary brain diseases.
  • cellular therapeutic agent refers to a medicinal product (US FDA regulation) used for the purpose of treatment, diagnosis, and prevention of cells and tissues prepared through isolation, culture, and special manipulation from humans. Or through a series of actions such as proliferating and screening living autologous, allogeneic, or heterologous cells in vitro or otherwise altering the biological properties of a cell to restore tissue function. Means the drug used for the purpose.
  • treatment means any action that improves or benefits the condition of the disease by administration of the cell therapy agent.
  • the route of administration of the cell therapy composition of the present invention may be administered via any general route as long as it can reach the desired tissue.
  • Parenteral administration for example, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration may be, but is not limited thereto.
  • composition may be formulated in a suitable form with a pharmaceutical carrier generally used for cell therapy.
  • a pharmaceutical carrier generally used for cell therapy.
  • 'Pharmaceutically acceptable' refers to a composition that is physiologically acceptable and does not cause an allergic or similar reaction, such as gastrointestinal disorders, dizziness or the like, when administered to a human.
  • Pharmaceutically acceptable carriers include, for example, water, suitable oils, saline, carriers for parenteral administration such as aqueous glucose and glycols, and the like, and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol.
  • Other pharmaceutically acceptable carriers may be referred to those described in the following references (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).
  • composition may also be administered by any device in which the cell therapy agent can migrate to the target cell.
  • the cell therapy composition of the present invention may include a therapeutically effective amount of cell therapy for the treatment of a disease.
  • therapeutically effective amount means an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as thought by a researcher, veterinarian, doctor or other clinician. This includes amounts that induce alleviation of the symptoms of the disease or disorder being treated.
  • the optimal cell therapy content can be readily determined by one skilled in the art and includes the type of disease, the severity of the disease, the amount of other components contained in the composition, the type of formulation, and the age, weight, general health, sex and diet of the patient. It can be adjusted according to various factors including the time of administration, the route of administration and the rate of secretion of the composition, the duration of treatment, and the drugs used simultaneously. In consideration of all the above factors, it is important to include an amount that can achieve the maximum effect in a minimum amount without side effects.
  • the daily dose of stem cells of the present invention is 1.0 ⁇ 10 4 to 1.0 ⁇ 10 10 cells / kg body weight, preferably 1.0 ⁇ 10 5 to 1.0 ⁇ 10 9 cells / kg body weight divided into one or several times May be administered.
  • the actual dosage of the active ingredient should be determined in light of several relevant factors such as the disease to be treated, the severity of the disease, the route of administration, the patient's weight, age and gender, and therefore the dosage may It does not limit the scope of the present invention in terms of aspects.
  • composition comprising the cell therapy of the present invention as an active ingredient in the treatment method of the present invention is rectal, intravenous (intravenous therapy, iv), intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular Or via the intradermal route.
  • intravenous therapy iv
  • intraarterial intraarterial
  • intraperitoneal intramuscular
  • intrasternal transdermal
  • topical intraocular Or via the intradermal route.
  • the present invention provides a method of treatment comprising administering to a mammal a therapeutically effective amount of said cell therapy composition of the invention.
  • mammal refers to a mammal that is the subject of treatment, observation or experiment, preferably human.
  • Fibroblast markers Thy1
  • neural stem cell markers nestin, sox1, sox2, pax6
  • MET or EMT markers snail. AFP, GATA4
  • gene expression patterns of markers related to maintaining chromosomal stability Zsacn4 were confirmed using quantitative PCR (FIG. 2).
  • the fibroblasts were transformed into neural stem cells using the culture medium under various conditions except one small molecule material from the conditions in which the eight small molecule materials identified in Example 1-1 were added all at once. It was. 1 ⁇ 10 5 human fibroblasts were prepared in a 60 mm dish, and the morphological changes of the cells were confirmed by changing the culture medium under various conditions every 2 to 3 days from the following day. In addition, changes in expression patterns of various genes were observed through the cells obtained during subculture.
  • the neural stem cells induced through the culture medium containing 6 or 8 small molecule compounds maintained the shape of the neural stem cells even when cultured for a long time with continuous passage.
  • PCR and immunocytochemistry were performed to determine whether neural stem cells express markers of neural stem cells.
  • neural stem cells express nestin through immunocytochemistry (ICC) staining, and PCR expression of the neural stem cell markers nestin, sox1, and musashi1 is similar to neural stem cells derived from human embryonic stem cells. It was confirmed that the degree (Fig. 4).
  • the morphology of the cells was observed at each of the early and late stages of induced neural stem cells.
  • the expression of nestin through immunocytochemistry (ICC) staining and gene expression of the nestin, sox1, and musashi1 neural stem cell markers using PCR were confirmed (FIG. 5).
  • chromosomal analysis confirmed the presence or absence of chromosomal abnormalities in the early and late stages of the induced neural stem cells (Fig. 6), compared with the growth rate of the cells during the proliferation and maintenance of neural stem cells derived from human embryonic stem cells ( 7).
  • the induced neural stem cells were found to be able to continue culturing without chromosomal abnormalities while maintaining the shape and properties of neural stem cells even during long-term culture.
  • bisulfite PCR showed methylation / acetylation of the promoter region of nestin, a neuronal stem cell marker, to confirm epigenetic genetic changes. It was confirmed that the characteristics of fibroblasts were completely changed to the characteristics of neural stem cells (Fig. 8).
  • Example 4 In vitro differentiation of induced human neural stem cells
  • Examples 1 to 3 confirmed that inducing neural stem cells from human fibroblasts using small molecule materials was successfully performed. More specifically, in order to confirm the characteristics and availability of neural stem cells, differentiation ability in vitro was confirmed. First, they differentiated into trigeminal neurons (astrocyte), oligodendrocyte (neuron) and neurons (neuron).
  • human fibroblasts are highly useful as cell therapeutics for the treatment of brain diseases.
  • Example 5 In vivo differentiation of induced human neural stem cells
  • Examples 3 and 4 it was confirmed in vitro that the neural stem cells derived from human fibroblasts can be successfully differentiated into trigeminal nerve cells using small molecule materials.
  • human neural stem cells induced in the brain of mouse (Oriental Bio, Balb / c) were transplanted. It was. Then, the mice transplanted with human neural stem cells induced for 3 to 7 months were continuously observed.
  • the brain of the mouse transplanted with the neural stem cells induced by the small molecule compound showed no tumor formation externally for 3 to 7 months (FIG. 11A), and brain tissue was confirmed by H & E (Haematoxylin and Eosin) staining. It was also confirmed that no tumor formation was observed (FIG. 11B).
  • the induced neural stem cells were successfully transplanted through the expression of neural stem cell markers (FIG. 11C), and the induced neural stem cells 3 to 7 months after transplantation expressed human cell-specific markers and neuron-specific markers. It was confirmed that the transplanted cells were differentiated into neurons (FIG. 11D). This indicates that human neural stem cells induced by small molecule compounds can be differentiated into neurons without transplantation after transplantation.
  • induced neural stem cells can be used as an optimal model cell to study the development of neurons and the pathogenesis of brain disease, and as a cell therapy for the treatment of brain diseases in humans due to the advantage of not generating tumors. The probability is very high.
  • the method of directly converting human fibroblasts into neural stem cells using only small molecule materials without introducing the genes according to the present invention secures a sufficient amount of cells and various kinds of functional neurons for use in cell therapy through induction of genetically stable neural stem cells. Because it can be differentiated and does not generate tumors, it is useful for brain disease cell therapy.

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Abstract

La présente invention concerne un procédé permettant la conversion de fibroblastes humains en cellules souches neurales et, plus spécifiquement, un procédé permettant la conversion directe de fibroblastes humains en cellules souches neurales uniquement à l'aide d'une combinaison de composés à petites molécules sans l'introduction de gènes étrangers; et une utilisation de ceux-ci. Le procédé selon la présente invention permettant la conversion directe de fibroblastes humains en cellules souches neurales à l'aide uniquement de matériels à petites molécules sans l'introduction de gènes est utile dans un agent thérapeutique cellulaire de maladies du cerveau, puisqu'il est possible de fixer une quantité suffisante de cellules à utiliser dans une thérapie cellulaire par l'induction de cellules souches neurales génétiquement stables, et de différencier en divers types de cellules nerveuses fonctionnelles, et aucune tumeur n'est provoquée.
PCT/KR2016/003819 2015-04-13 2016-04-12 Procédé de conversion directe de fibroblastes humaines en cellules souches neurales à l'aide de composés à petites molécules WO2016167528A1 (fr)

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US15/545,314 US10711245B2 (en) 2015-04-13 2016-04-12 Direct conversion method of human fibroblasts into neural stem cells using small molecules
CN201680019389.1A CN107454913B (zh) 2015-04-13 2016-04-12 使用小分子的人成纤维细胞至神经干细胞的直接转化方法
JP2017538402A JP6599468B2 (ja) 2015-04-13 2016-04-12 小分子化合物を利用したヒト線維芽細胞を神経幹細胞に直接転換する方法

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3321354A4 (fr) * 2015-05-19 2018-11-14 Stemlab Inc. Procédé pour induire des cellules précurseurs d'oligodendrocytes à partir de cellules somatiques humaines induites par oct4 par reprogrammation directe
CN112384613A (zh) * 2018-07-10 2021-02-19 株式会社片冈制作所 神经样细胞的制备方法
KR102637401B1 (ko) * 2022-09-05 2024-02-16 주식회사 스마트셀랩 레노그라스팀에 의한 줄기세포의 운동신경전구세포로의 유도 및 운동신경세포로의 분화

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